Photonic fence

ABSTRACT

A system for tracking airborne organisms includes an imager, a backlight source (such as a retroreflective surface) in view of the imager, and a processor configured to analyze one or more images captured by the processor to identify a biological property of an organism.

If an Application Data Sheet (ADS) has been filed on the filing date ofthis application, it is incorporated by reference herein. Anyapplications claimed on the ADS for priority under 35 U.S.C. §§ 119,120, 121, or 365(c), and any and all parent, grandparent,great-grandparent, etc. applications of such applications, are alsoincorporated by reference, including any priority claims made in thoseapplications and any material incorporated by reference, to the extentsuch subject matter is not inconsistent herewith.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims the benefit of the earliest availableeffective filing date(s) from the following listed application(s) (the“Priority Applications”), if any, listed below (e.g., claims earliestavailable priority dates for other than provisional patent applicationsor claims benefits under 35 USC § 119(e) for provisional patentapplications, for any and all parent, grandparent, great-grandparent,etc. applications of the Priority Application(s)).

PRIORITY APPLICATIONS

The present application constitutes a divisional of U.S. patentapplication Ser. No. 14/255,119, entitled PHOTONIC FENCE, naming asinventors, Roderick A. Hyde, 3ric Johanson, Jordin T. Kare, ArtyomMakagon, Emma Rae Mullen, Nathan P. Myhrvold, Thomas J. Nugent, Jr.,Nathan John Pegram, Nels R. Peterson, Philip Rutschman and Lowell L.Wood, Jr. filed Apr. 17, 2014, which is currently co-pending or is anapplication of which a currently co-pending application is entitled tothe benefit of the filing date.

The present application constitutes a continuation-in-part of U.S.patent application Ser. No. 12/657,281, entitled PHOTONIC FENCE, namingRoderick A. Hyde, 3ric Johanson, Jordin T. Kare, Nathan P. Myhrvold,Thomas J. Nugent, Jr., Nels R. Peterson, Lowell L. Wood, Jr. asinventors, filed Jan. 15, 2010, which is currently co-pending or is anapplication of which a currently co-pending application is entitled tothe benefit of the filing date, and which claims benefit of priority ofU.S. Provisional Patent Application No. 61/205,430, entitled PHOTONICFENCE, naming Roderick A. Hyde, 3ric Johanson, Jordin T. Kare, Nathan P.Myhrvold, Thomas J. Nugent, Jr., Nels R. Peterson, Lowell L. Wood Jr. asinventors, filed Jan. 15, 2009.

If the listings of applications provided above are inconsistent with thelistings provided via an ADS, it is the intent of the Applicant to claimpriority to each application that appears in the DomesticBenefit/National Stage Information section of the ADS and to eachapplication that appears in the Priority Applications section of thisapplication.

All subject matter of the Priority Applications and of any and allapplications related to the Priority Applications by priority claims(directly or indirectly), including any priority claims made and subjectmatter incorporated by reference therein as of the filing date of theinstant application, is incorporated herein by reference to the extentsuch subject matter is not inconsistent herewith.

SUMMARY

In one aspect, a system for tracking airborne organisms includes animager (e.g., a camera or scanner), a backlight source (e.g., aretroreflector), and a processor. The processor is configured to analyzeone or more images captured by the imager including at least a portionof the backlight source and to identify a biological property (e.g.,genus, species, sex, mating status, gravidity, feeding status, age, orhealth status) of an organism (e.g., an insect, such as a mosquito, abee, a locust, or a moth) in the field of view of the imager, usingcharacteristic frequency, harmonic amplitude, shape, size, airspeed,ground speed, or location. The system may further include anillumination light source arranged to illuminate the field of view ofthe imager. The organism may have wings, in which case the processor maybe configured to identify the biological property using a wingbeatfrequency.

The system may further include a detector configured to detect a signalindicative of a property of an organism in the field of view of theimager. For example, the detector may include a photodiode, which may beconfigured to detect light from an optional targeting light sourceconfigured to be directed at the organism, or light from the backlightsource. The targeting light source may be configured to be directed atthe organism from a plurality of directions (e.g., a group of spotlightsor LEDs which may be placed at positions surrounding an expectedorganism location). The detector may be configured to detect a signalindicative of a distance from the imager to the organism, for example bydetecting shadows cast by the organism in a plurality of targeting lightsources (which may, for example, be different colors or be configured tobe selectively switched on and off), or by using a plurality of opticalposition sensing devices to triangulate the organism. The processor (ora second processor) may be configured to use this signal to determine adistance from the imager to the organism. Alternatively, the processormay use one or more images captured by the imager to determine thedistance to the organism, for example in cases where the imager includesa plurality of imaging devices, which may function in the same ways asthe targeting light sources described above. The detector may have abandwidth greater than one-half of a frame rate of the imager, or lessthan or equal to a frame rate of the imager, and may have an imageresolution or field of view greater or smaller than that of the imager.The detector may also be acoustic.

In another aspect, a method of tracking airborne organisms includesacquiring a first image from an imager, the imager having a backlightsource (e.g., a retroreflector) in its field of view, determining thatthe image includes an organism at a location, acquiring a second image,and determining a biological property (e.g., genus, species, sex, matingstatus, gravidity, feeding status, age, or health status) of theorganism using the second image (e.g., by determining characteristicfrequency, harmonic amplitude, shape, size, airspeed, ground speed,flight direction, flight path, or location). The first and second imageshave different resolutions (e.g., the first image may be finer orcoarser than the second image), or they are acquired at different framerates (e.g., the second image may be acquired at a faster or slowerframe rate than the first). The images may also differ in size.Acquiring the first or second image may include illuminating the regionof the acquired image, for example with a laser or an LED. Acquiringeither image may include acquiring a series of images. The images mayboth be acquired by the imager, or the second image may be acquired by adifferent device (e.g., a photodiode).

In another aspect, a system for disabling airborne organisms includes animager (e.g., a camera or scanner), a backlight source (e.g., aretroreflector), a processor, and a disabling system. The processor isconfigured to analyze one or more images captured by the imagerincluding at least a portion of the backlight source and to identify abiological property (e.g., genus, species, sex, mating status,gravidity, feeding status, age, or health status) of an organism (e.g.,an insect, such as a mosquito, a bee, a locust, or a moth) in the fieldof view of the imager, using characteristic frequency, harmonicamplitude, shape, size, airspeed, ground speed, or location. Thedisabling system is configured to disable the organism (e.g., bykilling, damaging a wing or antenna, or impairing a biological function)responsive to the identified property (e.g., only disabling organisms ofa determined genus, species, sex, or gravidity). The disabling systemmay include a laser (e.g., a UV-C laser or an infrared laser), and maybe configured to accept location data from the processor for use intargeting the organism.

In another aspect, a method of disabling airborne organisms includesacquiring a first image from an imager, the imager having a backlightsource (e.g., a retroreflector) in its field of view, determining thatthe image includes an organism at a location, acquiring a second image,determining a biological property (e.g., genus, species, sex, matingstatus, gravidity, feeding status, age, or health status) of theorganism using the second image (e.g., by determining characteristicfrequency, harmonic amplitude, shape, size, airspeed, ground speed,flight direction, flight path, or location), and disabling the organismresponsive to the determined biological property (e.g., killing theorganism or impairing a body function such as mating, feeding, flying,hearing, acoustic sensing, chemosensing, or seeing). The first andsecond images have different resolutions (e.g., the first image may befiner or coarser than the second image), or they are acquired atdifferent frame rates (e.g., the second image may be acquired at afaster or slower frame rate than the first). The organism may bedisabled, for example, by directing a laser beam at the organism(optionally using targeting information obtained from one or both of theacquired images), by directing an acoustic pulse at the organism, byreleasing a chemical agent, or by directing a physical countermeasure atthe organism.

In another aspect, a system for identifying status of flying insects ina region includes an imager, a backlight source (e.g., a retroreflector)configured to be placed in the field of view of the imager, and aprocessor configured to analyze one or more images captured by theimager including at least a portion of the backlight source, theprocessor being configured to identify probable biological status of aninsect in the field of view of the imager using characteristicfrequency, shape, size, airspeed, ground speed, or location. The insectmay be a mosquito, in which case the processor may be configured todetermine a probability that the mosquito is infected with malaria. Theprocessor may be configured to gather probable biological status of aplurality of insects, for example gathering population data for apopulation of insects, or gathering probable biological status data as afunction of an environmental parameter (e.g., time of day, season,weather, or temperature).

In another aspect, a system for tracking airborne organisms includes animager, a backlight source (e.g., a retroreflector) configured to beplaced in the field of view of the imager, a processor, and a detectorconfigured to detect an organism in the field of view of the imager. Atleast one of the imager and the detector is configured to collect colordata. The processor is configured to analyze one or more images capturedby the imager including at least a portion of the backlight source, andto identify a biological property of an organism in the field of view ofthe imager using at least one datum selected from the group consistingof characteristic frequency, harmonic amplitude, shape, size, airspeed,ground speed, and location. The system may use the collected color datato determine a probable engorgement status of the organism (e.g., amosquito engorged with blood). The system may further include aforward-facing light source configured to illuminate the organism, forexample when it is in the field of view of the imager or of thedetector. The detector may include a photodiode (e.g., a quad cellphotodiode). The system may further include a targeting light sourceconfigured to be directed at the organism from one or more directions,in which case the photodiode may be configured to detect light reflectedfrom the organism or light from the backlight source. The detector maybe configured to detect a signal indicative of a distance from theimager to the organism. The processor (or a second processor) may beconfigure to determine a distance from the imager to the organism usingthe signal detected by the detector. The processor may be configured todetermine a distance from the imager to the organism by using the signaldetected by the detector. The system may include a plurality oftargeting light sources in differing positions (e.g., different coloredlight sources), so that the detector may detect shadows cast by theorganism in each light source. These targeting light sources may beconfigured to be selectively switched on and off. The detector mayinclude a plurality of optical position sensing devices configured toprovide range information by triangulation of the organism. The detectormay have a bandwidth greater than one-half the frame rate of the imager,or of less than or equal to the frame rate of the imager, and may havean image resolution that is less than or greater than the imageresolution of the imager. The processor may be configured to identifygenus, species, sex, age, mating status, gravidity, feeding status, orhealth status of the organism. The system may further include adisabling system responsive to the identified property configured todisable the organism.

In another aspect, a method of tracking airborne organisms includesacquiring a first image (e.g., a monochrome or a color image) having afirst image resolution from an imager with a backlight source (e.g., aretroreflector) in its field of view, determining that the imageincludes an organism at a location, acquiring a second image having asecond image resolution and including color data (e.g., with aphotodiode such as a quad cell photodiode or with an imager), anddetermining a biological property of the organism (e.g., genus, species,sex, mating status, gravidity, feeding status, age, or health status)using at least the second image, where the first and second imagesdiffer in resolution or frame rate, or the second image includes colordata not included in the first image. Determining the biologicalproperty (e.g., engorgement status) may include using the color data,and may include determining characteristic frequency, harmonicamplitude, shape, size, airspeed, ground speed, flight direction, flightpath, or location.

In another aspect, a system for tracking airborne organisms includes animager, a backlight source (e.g., a retroreflector) configured to beplaced in the field of view of the imager, and a processor configured toanalyze one or more images captured by the imager, the processor beingconfigured to identify a rotation of an organism in the field of view ofthe imager. The processor may be configured to determine a revolutionrate of the organism, and may further be configured to determine awingbeat frequency of an organism that has wings. The system may furtherinclude a detector (e.g., a photodiode such as a quad cell photodiode)configured to detect a signal indicative of a property of an organismsin the field of view of the imager. The system may further include atargeting light source (from one or more directions), and the photodiodemay be configured to detect light from the light source reflected fromthe organism or light from the backlight source. The detector may beconfigured to detect a signal indicative of a distance from the imagerto the organism, for example to be determined by the processor or by asecond processor. The system may include a plurality of targeting lightsources at differing positions, where the detector is configured todetect shadows cast by the organism in each light source, or thedetector may include a plurality of optical position sensing devicesconfigured to provide range information by triangulation of theorganism. The detector may have a bandwidth greater than about one-halfof a frame rate of the imager, or of less than or about equal to a framerate of the imager, and may have an image resolution less than orgreater than the image resolution of the imager. The processor may beconfigured to identify a biological property of the organism selectedfrom the group consisting of genus, species, sex, mating status,gravidity, feeding status, age, and health status. The system mayfurther include a disabling system configured to disable the organism.

In another aspect, a method of tracking airborne organisms includesacquiring a first image from an imager having a backlight source (e.g.,a retroreflector) in its field of view, determining at the imageincludes an organism at a location, and determining that the organism isrotating about a revolution axis. The method may further includedetermining a revolution rate or revolution axis for the organisms, ordetermining a wingbeat frequency for an organism with wings. It mayinclude determining a biological property of the organism (e.g., genus,species, sex, mating status, gravidity, feeding status, age, or healthstatus), which may include determining a datum selected from the groupconsisting of characteristic frequency, harmonic amplitude, shape, size,airspeed, ground speed, flight direction, flight path, and location, andmay include responding to the determined biological property bydisabling the organism. The method may further include detecting asignal indicative of a distance from the imager to the organism.

In another aspect, a system for tracking organisms includes an imager, abacklight source (e.g., a retroreflector) configured to be placed in thefield of view of the imager, a processing configured to analyze one ormore images captured by the imager and to identify a biological property(e.g., genus, species, sex, mating status, gravidity, feeding status,age, or health status) of an airborne organism (e.g., an insect such asa mosquito or a psyllid) in the field of view of the imager, and aphysical trap configured to physically capture at least one organism(e.g., a flying organism, or an immature individual of a species that iscapable of flight at maturity), wherein the system is configured to usethe identified biological property to measure an efficacy of thephysical trap. Measuring the efficacy of the physical trap may includecomparing a number of organisms in the trap with a number of airborneorganisms identified by the processor (e.g., during the same timeinterval or during a different time interval). The field of view of theimager may include at least a portion of the trap interior, or it mayinclude a volume exterior to the trap.

In another aspect, a method of determining efficacy of a trap forairborne organisms includes monitoring a population of airborneorganisms to determine a population in a monitored space by acquiring animage from an imager having a field of view including the monitoredspace and a backlight (e.g., a retroreflector), determining that theimage includes an organism (e.g., an insect such as a mosquito or apsyllid), and determining a biological property of the organism (e.g.,genus, species, sex, mating status, gravidity, feeding status, age, orhealth status), determining a number of airborne organisms captured by atrap, and comparing the number of captured organisms with the determinedpopulation of airborne organisms. Comparing the number of capturedorganisms with the determined population of organisms may includecomparing only organisms having a selected biological property, orcomparing a fraction of organisms having a selected biological property.The trap may be configured to capture flying organisms, or immatureindividuals of a species that is capable of flight at maturity.

In another aspect, a system for tracking airborne organisms includes aphysical trap configured to capture at least one airborne organism(e.g., an insect such as a mosquito or a psyllid), a detection componentconfigured to identify a biological property (e.g., genus, species, sex,mating status, gravidity, feeding status, age, or health status) of thecaptured organism, the detection component including an imager, abacklight source (e.g., a retroreflector) configured to be placed in thefield of view of the imager, and a processor configured to analyze oneor more detected images to identify the biological property, and anotification component configured to send a notification to a remoteuser in response to the identified property.

The foregoing summary is illustrative only and is not intended to be inany way limiting. In addition to the illustrative aspects, embodiments,and features described above, further aspects, embodiments, and featureswill become apparent by reference to the drawings and the followingdetailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a schematic of a detection system.

FIG. 2 illustrates an embodiment of a system surrounding a structure.

FIG. 3 is a control flow diagram for an implementation of a tracking anddosing system.

FIG. 4 is a photograph of a damaged mosquito wing.

FIG. 5 is a lethality graph for a series of mosquito IR laser exposures.

FIG. 6 is a lethality graph for UV laser exposures for femalemosquitoes.

FIG. 7 is a lethality graph for UV laser exposures for male mosquitoes.

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description, drawings, and claims are not meant to be limiting.Other embodiments may be utilized, and other changes may be made,without departing from the spirit or scope of the subject matterpresented here.

As shown in FIG. 1, a system for locating or identifying informationabout, or optionally disabling insects or other organisms includes animager 10, illumination source 12, a retroreflective surface 14, aprocessor 16 configured to analyze images captured by the imager 10, atargeting laser 18, and a photodiode 20. In the illustrated embodiment,imager 10 is a CMOS camera placed at the base of support post 22, but avariety of other imagers may be appropriate. For example, CCD-baseddetectors, scanning systems, or other types of detectors may beimplemented. Moreover, in some approaches two or more imagers may beplaced on support post 22 or on other supports. In some embodiments,retroreflective surface 14 may be replaced with a light emitting surface(backlight), for example a substantially uniform light emitting surfacewith a desired angular distribution at light, which may be aimed towardimager 10.

As illustrated, retroreflective surface 14 is placed on adjacent supportpost 24 spaced apart from the support post 22 to define an intermediateregion. In some embodiments, imagers or retroreflective surfaces may beplaced on multiple support posts. For example, in some embodiments,support posts may be arranged to surround an area of interest, asillustrated in FIG. 2, and imagers or retroreflective surfaces may bearranged on the support posts so as to view all, substantially all, orat least a portion of the entrances to the area of interest. Whileelements placed on support post 22 in FIG. 1 have been placed apart forclarity in illustration, in practice they may be more closely spaced.

In the illustrated embodiment, support posts 22 and 24 have a heightselected to exceed the typical flying height of an insect of interest.For example, more than 99% of Anopheles mosquitoes (which may carrystrains of malaria that can infect humans) fly at less than 3-5 metersof altitude, so support posts of 3-5 meters may be used in a system thatcan view substantially all mosquitoes passing through an area ofinterest. The width of support posts 22 and 24 is selected to provideadequate support and surface area for components includingretroreflective surface 14; in the illustrated embodiment, the supportposts are 10-20 cm wide, and are placed 100 m apart. The width ofretroreflective surface 14, and of the field of view of imager 10, maybe selected as a function of the flight speed of the target(s) ofinterest and the frame rate of imager 10, such that the silhouette of aninsect will be within the field of view for at least one full frameinterval, and as a function of the flight speed and the desired wingbeatsensing accuracy, such that the silhouette will be within the field ofview for a sufficient period to make a measurement of the desiredaccuracy.

Illumination source 12 (which may be, for example, a laser, an LED, anincandescent light, a mirror reflecting sunlight, or any other suitablelight source) directs light from support post 22 toward support post 24to illuminate the retroreflective surface 14 on support post 24. In theillustrated embodiment, illumination source 12 is an LED producing afan-shaped beam. Retroreflector 14 returns light to imager 10. When anorganism 26 (such as a mosquito) travels between posts 22 and 24, theorganism appears as a dark shadow on the retroreflective background 14or as a break in a beam of light. Upon detecting such a shadow, in someembodiments, imager 10 may shift to a higher frame rate or a higherspatial resolution local to the shadow. Alternatively, a second imager(not shown) may be employed to collect a higher frame rate or higherresolution image in a small region local to the shadow. The higher framerate image may be used, for example, by processor 16 to identify awingbeat frequency for the mosquito (or other flying organism). In someembodiments, the sensing of organism 26 may trigger a forward-facinglight. In other embodiments, a forward-facing light may be always on, orturned on when ambient light is low. Forward-facing illumination isexpected to be preferred if it is desired to identify color data for theorganism. In some embodiments, forward-facing light may be provided bytargeting laser 18, or by a more broad-band source (not shown). Wingbeatfrequency and harmonics may be used to determine probable species, sex,and other biological properties such as mating status of a mosquito; forsome information on characteristic frequencies, see Robertson, et al.,“Heritability of wing-beat frequency in Anopheles quadrimaculatus,” J.Amer. Mosquito Control Assoc., 18(4):316-320 (2002); Moore, “ArtificialNeural Network Trained to Identify Mosquitoes in Flight,” J. InsectBehavior, 4(3):391-396 (2005); “An Automated Flying-Insect DetectionSystem,” NASA Technical Briefs, SSC-00192 (2007), available at<www.techbriefs.com/content/view/2187/34/>; Göpfert, et al.,“Nanometer-range acoustic sensitivity in male and female mosquitoes,”Proc. Biol. Sci. 267(1442):453-457 (2000); and Gibson, et al., “Flyingin Tune: Sexual Recognition in Mosquitoes,” Curr. Biol. 16:1311-1316(2006), all of which are incorporated herein by reference.

In some embodiments, periodic data which is not directly related towingbeats may be collected. In particular, it has been observed thatAsian Citrus Psyllids rotate in space as they launch themselves into theair, and these rotations have a periodicity that may be captured by asystem such as that shown in FIG. 1. See, e.g., our video available at<www.youtube.com/watch?v=fMu8n1_8Ozg>. In some embodiments, processor 16may be configured to identify such rotation and separate it fromwingbeats using data from imager 10. In such embodiments, higher framerate and/or a second imager as described above may have utility inidentifying rotary movements of the organism. In some embodiments, suchrotary movements may be used to identify a species or other biologicalproperty of the organisms.

In some embodiments, harmonic frequency spectra may be of significantutility in identifying mosquitoes or other insects. For example, thesecond harmonic frequency of the wingbeats of certain honeybee speciesare substantially similar to the wingbeat frequency of certain speciesof mosquitoes. Thus, in some embodiments, spectral analysis of harmonicfrequencies may be used to prevent spurious identification of honeybeesas mosquitoes. In addition, concentrating on higher-frequency harmonicsmay allow faster detection and identification of insects in someembodiments by reducing the time period necessary to identify thefrequencies present. Chen, et. al, have described a system using suchspectra to identify mosquitoes and other insects. See Chen, et al.,“Flying Insect Classification with Inexpensive Sensors,” published at<arxiv.org/pdf/1403.2654v1>, a copy of which is included herewith andincorporated by reference herein.

In some embodiments, processor 16 may incorporate a graphics processingunit (graphics card) for analysis. The graphics processing unit (GPU)may have a parallel “many-core” architecture, each core capable ofrunning many threads (e.g., thousands of threads) simultaneously. Insuch a system, full-frame object recognition may be substantiallyspeeded as compared to traditional processors (e.g., 30 times as fast).In some embodiments, a field-programmable gate array may be directlyconnected to a high-speed CMOS sensor for fast recognition.

In addition to the higher-speed camera imaging of the organism, thesystem may also employ a targeting laser 18 (or other suitable nonlaserlight source) and detector (such as photodiode 20) to confirmcharacteristics of organism 26. For example, if processor 16 identifiesa morphology or frequency suggestive of an organism of interest (such asa mosquito), targeting laser 18 may be directed at organism 26 usinglocation information from processor 16. The reflection of targetinglaser 18 from organism 26 is detected by photodiode 20. In someembodiments, this reflection may have relatively lower image resolutionbut a very fast frame rate, wide frequency response, or a highsensitivity to changes in cross section of the organism. The signal fromthe photodiode may be used, for example, to measure wingbeat frequencyor harmonics very accurately to identify the organism or to otherwiseclassify the organism into an appropriate category, or otherwisedistinguish the organism. Targeting laser 18 may also or alternativelyprovide additional light for higher frame rate or higher resolutionimage acquisition by imager 10.

The second imager or targeting laser 18 may be aimed by a galvanometer,MEMS device, or other suitable optical pointing systems. In someembodiments, the second imager or targeting laser 18 may be aimable intwo dimensions, while in others, a single-axis galvanometer system maybe used to allow the targeting laser to track within a single firingplane. In one-dimensional systems, a series of two-dimensional imagescaptured by imager 10 may be used to predict when organism 26 will crossthe firing plane, at which point it may be illuminated by targetinglaser 18. In some embodiments, targeting laser 18 may be continuouslyscanned through space, for example by a rotating or oscillating mirror,and fired when its projected path intersects with the organism. In somesuch embodiments, the scan path may be dynamically adjusted, for exampleto provide a dwell time at a target location.

While the targeting laser 18 is described as being aimed by agalvanometer, MEMS device, or other targeting system, such aiming may beimplemented via direct physical positioning of the laser, or throughdirection by an optical system, including conventional opticalcomponents, such as acoustical optic scanners, scanning mirrors, orsimilar. In some embodiments, a phase detection autofocus system such asthat described in U.S. Pat. No. 6,480,266, which is incorporated byreference herein, may be used to focus the laser at the point ofinterest.

In some embodiments, once the organism has been identified or otherwisecategorized or characterized, it may be desirable to take action todisable or destroy the organism. For example, in some embodiments, whena mosquito has been detected as entering the field of view, acountermeasure such as a laser beam may be used to disable or destroythe mosquito. In such embodiments, location information for the organism26 may be passed from the imager 10, the processor 16, the targetinglaser 18, or an associated targeting processor, not shown, to a dosinglaser 28. In some embodiments, other countermeasures might include asonic countermeasure transmitted by an acoustic transducer, a physicalcountermeasure such as a solid or liquid projectile, or a chemicalresponse, in lieu of or in addition to dosing laser 28. In someembodiments, targeting laser 18 and dosing laser 28 may be the samecomponent, for example using a higher amplitude for dosing than fortargeting. In other embodiments, targeting laser 18 and dosing laser 28may be separate components. In this case, they may optionally use acommon aiming and/or focusing mechanism such as a beam splitter or beamcombiner that allows dosing laser 28 to fire along the same path astargeting laser 18. FIG. 3 is a control flow diagram for animplementation of the tracking and dosing system, illustratingcooperation of imager assembly 40, processor 42, targeting laserassembly 44 and dosing laser assembly 46.

In some embodiments, undesirable organism 26 may be killed by dosinglaser 28. In other embodiments, dosing laser 28 may instead disableorganism 26 in a variety of ways. For example, if it is desired toinhibit spread of malaria, it may be sufficient to impede a femalemosquito's ability to blood feed, disrupting the disease cycle. In someembodiments, this may be accomplished by damaging or destroying theantennae. Damage to the antennae may also inhibit mating behavior, whichmay reduce the overall mosquito population if enough mosquitoes in aregion can be dosed. In some embodiments, reproduction may also beslowed or prevented by impairing fertility of the female or the malemosquito. Radiative treatment may also impair the metabolic efficiencyof mosquitoes or other insects, or may damage essential body structuressuch as the wings or eyes without immediately killing the insect. FIG. 4is a photo of a mosquito wing which has been damaged by laser treatment.

In some embodiments, rather than or in addition to targeting organismsfor destruction, the system of FIG. 1 may be used as a census-takingdevice. If desired, the system may be left unattended for a substantialperiod of time to determine activity as a function of time of day,weather, season, or other changing environmental parameters, and flightcharacteristics of different organisms may be tracked over time. Byanalyzing shape, size, wingbeat frequency, wingbeat harmonics, position,flight patterns, airspeed, or groundspeed, information about biologicalproperties such as genus, species, gender ratios, age distribution,mating status, and the like may be determined for the organismpopulation. In some embodiments, it may be possible to determinedisease-carrying status, since it is expected that disease carriers suchas malarial mosquitoes will have different characteristics perceivableby the system (e.g., flight characteristics, shape, size) due to bodystresses associated with illness. In some embodiments, thesecharacteristics of disease-carrying organisms may be identifiable viastatistical bias (e.g., while the system may not identify individuals asdiseased, it may be able to tell that some fraction of the individualsobserved are diseased). Such embodiments may be useful for targetingdisease mitigation strategies into areas of highest infection rate, forexample. In embodiments including a dosing laser or othercountermeasure, in circumstances where it is undesirable or impracticalto incapacitate all mosquitoes (or other insect pests), discriminationby sex or other biological status may allow more effective eradicationof the population as a whole (for example, by preferentially targetinggravid females, females ready for mating, or mosquitoes already infectedwith malaria). In some embodiments, identification of a particularbiological property (of an individual or a population) may trigger anotification to be sent to a remote location. For example, if a singleAsian Citrus Psyllid is detected in an area expected to be free of them(e.g., an orchard), the system may notify the farmer (or any appropriateremote user) so that countermeasures can be taken and defenses examinedfor “leaks,” or if the system identifies a noticeable increase inpopulation of mosquitoes or of malarial mosquitoes in a particularregion, it may notify doctors and/or scientists so that the change canbe promptly addressed.

While the embodiments described herein have related to ground-basedsystems mounted upon fixed vertical supports, a variety of other designconfigurations may be implemented by one of skill in the art. In someembodiments, a substantial portion of the components or even all of thecomponents may be mounted upon a single support unit. For example, asingle post having lasers and cameras at the top may illuminate and viewa surrounding horizontal ring of retroreflector, forming a conical ortent-like detection area. For another example, one or more lasers andcameras may be rotated or translated so as to sweep the narrow camerafield of view across a large volume, so as to detect insects anywherewithin a volume (such as a room); in this case a large area ofretroreflector material such as a retroreflective paint or tape can beapplied to one or more walls of the room.

In one approach, one or more components may be mounted on a movingsupport such as a ground-based vehicle, air-based vehicle (e.g., a UAV),or other vehicle. If imager and targeting or dosing lasers are mountedon an airborne vehicle, it may be impractical to provide aretroreflective surface as described above. In some such embodiments,organisms may be located by ground-looking radar. For a vehicletraveling at 50 m/s and scanning a 100 m swath of ground, a relativelymodest transmitter power (in the tens to hundreds of milliwatts) mayprovide an adequate resolution for locating organisms for a targetinglaser.

In some embodiments, the imager or the detector may receive light thatis produced responsive to the illuminating light. For example, asdescribed in Belisle, et al., “Sensitive Detection of Malaria Infectionby Third Harmonic Generation Imaging,” Biophys. J. 94(4):L26-L28 (2008),which is incorporated herein by reference, certain components of tissueor residue such as biological waste products (e.g., hemozoin crystalsproduced by malarial mosquitoes) may produce wavelengths of lightdifferent from the illuminating light through any of a variety effects,including three photon effects. In one such approach, illuminating lightmay be selected to correspond to a response of hemozoin. The detectormay then detect light at a frequency corresponding to a resonance of thehemozoin.

It may be appropriate in some applications to provide a guard regionaround the targeting or dosing light beam. In such an approach, anappropriate detection system may determine the presence of objects ororganisms within a region surrounding the target object. If such anobject or organism is detected, the system may determine that it isinappropriate to activate the targeting or dosing light source, forexample, to prevent damage to such objects or organisms. In one example,the guard region may be configured to detect the presence of humans ordomestic animals within a selected proximity of the area to beilluminated. Such systems may be implemented using the illuminatinglight source, or an alternative light source, such as an LED or similarsource arranged to illuminate a region surrounding the expected path ofthe targeting or dosing beam. Alternatively, the imaging system maydetect humans or domestic animals in the field of view and avoidtransmitting the targeting or dosing light beam.

In some cases the illuminating light source may have sufficient power tocause harm, for example if a person or animal looks directly into thelight source. The system may be configured to detect the presence oflarge obstructions and turn off or reduce the power of the illuminatinglight source before harm is done.

It will be understood that “identification” of organisms (such asmosquitoes and other insects) on the basis of wingbeat characteristics,morphology, or other measurements, may be probabilistic in nature. Forexample, it may be determined that it is more likely than not that agiven organism is a gravid female Anopheles mosquito, and actions may betaken on that probability, even though other genera, sexes, or statusescannot be ruled out.

Maintenance and Olfactory Testing of Mosquito Population

We have maintained and tested a population of Anopheles stephensi in aninsectary. The mosquitoes were kept in a maintained environment of a 12h:12 h light:dark cycle; air temperature 80° F.±10° F. and 80%±10%humidity. Adult mosquitoes were held in a variety of containers.Breeding populations were placed into 12″×12″ white semi-transparentplastic containers with plastic mesh sides and a front sleeve for easyaccess. To sugar feed the adults, we used a Petri dish full of raisins.We placed a Petri dish lined with 9 cm filter paper, filled with waterinside the cage. This dish functioned as water source as well as an egglaying cup. The bottom of the cage was covered with absorbent papertowel to limit fungal growth due to urine and blood excretions of thefemales.

When adult mosquitoes were about six to ten days old, we blood fed thefemales while they were still inside their cages. We used Hemostat brandsheep blood. The feeding apparatus was a 10 cm Plexiglass Petri dishwhich had a copper coil tube glued to the bottom and circulated warmwater to keep the blood at body temperature. The bottom of the feedingapparatus was filled with water at 98° F. We stretched parafilm toloosely cover the water in Petri dish. Then sheep blood was added to theapparatus and another layer of Parafilm was stretched to cover theblood. A bucket of water between 98 and 100° F. was placed in theinsectary. It was hooked to the copper tubing of the feeding apparatususing plastic tubing and fittings. Inside the bucket there was anaquarium pump and a heater that circulated the warm water to the feedingapparatus. The feeding apparatus was placed in a cage through thesleeve. The sleeve was secured around the plastic tubing and mosquitoeswere allowed to feed until satiation. Once females had taken blood, theywere observed to find a quiet spot to rest and digest. Three to fivedays later eggs were laid in groups of 50 to 200 on the surface of thewater. These eggs hatched after two days. (See, Benedict, M. Q., inMolecular Biology of insect disease vectors. Ed. Crampton, Beard andLouis. Chapman and Hall, London, pp. 3-12, 1997, which is incorporatedby reference herein)

Experimental cages, hereinafter referred to as cradle to grave (C2G)boxes, were made of 12″×12″ interconnected clear acrylic. The sides andbottoms of the boxes were glued together, and they were reinforced bytabs for additional security. For ease of cleaning and access duringmanipulation of the mosquitoes, the top of the cage was not glued intoplace. There were two 6″ diameter openings on opposite sides of thecage. The one in the front was covered with a sleeve and the one on theback was lined with fine mesh, providing a texture on which mosquitoescould land. On the front, 2.5″ to the right of the sleeve and 2″ below,there was a 0.5″ diameter pipe fitting covered with mesh. This fittingwas used to connect a CO₂ tank during anesthetization of mosquitoes.While the mosquitoes were anesthetized, the lid was removed andmosquitoes could be handled for experiments.

There are certain advantages of using a cradle to grave box over othertypes of mosquito containment cages. Cradle to Grave boxes are clear;they allow the experimenter to observe behavior or document data withoutobstructed view. Another advantage of the box over conventional cage islimiting the number of times mosquitoes are handled. 50 to 100 pupaewere placed in a Cradle to Grave box and allowed to emerge. Once theadult mosquitoes were four to five days old, they were ready forexperimental manipulations. The port made of 0.5″ pipe fitting can beattached to CO₂ for anesthetization; this eliminates the need to chillthe mosquitoes, and consequently condensation does not occur duringvarious methods of cold application. Our handling experiences suggestusing aspirators during mosquito retrieval may affect their lifespanadversely. In the Cradle to Grave box, there is typically no need toaspirate mosquitoes into other containers.

White plastic rectangular trays (15″×7″×1.5″) were used to containlarvae. Once the eggs were laid, they were washed carefully into a whitetray for hatching. To provide food for larvae, 50% w/w active (live)baker's or brewer's yeast and ground tropical fish flakes were added towhite trays. The trays are filled halfway with distilled water.Achieving the right density of larvae in trays is known to be importantin their growth and development. The most common problems associatedwith overcrowding are longer development time, reduced pupation andeclosion, and a decrease in pupal weight. Studies have shown thatcrowded larvae exhibit several negative effects: lower weight atemergence, quantity of the blood meal and lower overall fertility rates(Benedict, 1997). If trays are overcrowded, thinning the larvae ispreferred to maintain a healthy population. After the fourth molting,pupae develop. Pupae were collected daily and placed into the opaquebreeding cages for continuation of the colony, or transferred into clearexperimental cages.

Adult mosquitoes were retrieved from their cage into smaller containersusing an aspirator made of two clear tubes connected to an electricpump. These retrieval boxes were 3.5″×3.5″×2.5″ and made of clearacrylic. One side of the box had a 2.44″ diameter opening which iscovered with fine mesh and allowed air flow as well as providing atextured surface for mosquitoes. One side of the retrieval box had two0.5″ pipe fittings that were used to connect tubes. These pipe fittingscould be plugged with acrylic rods when the aspirator was not in use.

After mosquitoes were anesthetized with CO₂ for experimental purposes,fine camel brushes were also used to change the position of themosquitoes.

To identify and assess the olfactory behavior of mosquitoes, we designeda bioassay, based on an olfactometer similar to that described in Geieret al., Entomol. Exp. Appl. 92:9-19, 1999 (which is incorporated byreference herein; see also Braks, et al., Physiological Entomology26:142-148, 2001, incorporated by reference herein), which met thefollowing requirements:

-   -   1. Monitoring of all behavioral sequences in the host finding        process such as perception, activation, orientation towards the        odor source, and landing.    -   2. Simple and fast testing of many odor samples in a limited        time.    -   3. Easy comparison of extracts from natural odor sources or        synthetic attractants (see, e.g., Miller, et al., In Chemical        Ecology of Insects, W. J. Bell, & R. T. Cardé (eds.), Chapman        and Hall, New York, pp. 127-157, 1984; Sutcliffe, Insect Science        and its Application 8: 611-616, 1987, both of which are        incorporated herein by reference).    -   4. Wide measuring range to differentiate the strength of        attractive stimuli.    -   5. Easy clean-up to avoid contamination caused by previous        stimuli (Schreck, et al., J. Am. Mosquito Control Assoc. 6:        406-410, 1990, which is incorporated herein by reference).

The olfactometer was constructed out of 7 mm thick transparent acrylicsheets. Twelve Y-shaped layers were placed on the acrylic base andbolted together on a metal table. Screened removable chambers werelocated at each end: a release chamber at the base of the Y-shape, andtwo chambers at the end of the arms. A transparent removable lid wasbolted to the layers below and provided containment for mosquitoes. Theresulting construction allowed for easy observation during experiments.

A 12 V fan was attached to the release chamber providing a wind-tunneleffect, luring mosquitoes away from the stimulus. Mosquitoes traveled 89cm to reach the stimulus chambers.

In a standard experiment, at least 25 female mosquitoes were aspiratedinto the release chamber using the human hand as bait. This procedureensured that all mosquitoes used in the test were ready to seek for ahost. The release chamber was made of clear acrylic, which was sized3.22″×3.22″×3.24″. Two sides of the release chamber had acrylic screens,one of which was removable for cleaning or other manipulation purposes.The release chamber also had two 0.5″ pipe fittings to connect anaspirator or CO₂ source as needed.

Five minutes after the release chamber was attached to the olfactometer,the test stimulus was presented in one arm while the control chamberremained empty. At the same moment, the release chamber opened andmosquitoes entered the device. The fan was then turned on to lure themosquitoes back into the release chamber. Five minutes into theexperiment, the mosquitoes were counted (those mosquitoes remaining inthe release-, stimulus-, and control chambers, respectively). At the endof the experiment, CO₂ gas was pumped through the stimulus chambers andanesthetized mosquitoes transferred back to the insectary.

Olfaction experiments such as those described herein may be used to testattractants for bringing species within range of the targeting system.They may also be used to determine whether mosquitoes' ability to seekhuman prey has been affected by dosing with photons as described herein.

Mosquito Vulnerabilities

In general, nocturnally active blood-feeding mosquitoes such as theAfrican malaria mosquito Anopheles gambiae locate and identify theirvertebrate hosts primarily by odor. The olfactory organs in adult femalemosquitoes are associated with the antennae and maxillary palp. Theseare covered by hair-like sensilla. The sensilla are innervated byolfactory receptor neurons as well as by mechano-, thermo-, orhygroreceptor cells. The olfactory cues exhaled in the breath (e.g.,carbon dioxide) or excreted from the skin (e.g., components of sweat)are detected by the sensilla, allowing the female mosquito to home in ona potential human host. (See, e.g., Ghaninia et al., Eur J Neurosci.26:1611-1623, 2007). The dependence upon the antennae and maxillary palpfor sensing the proximity of a human host suggests that disruption ofthese important sensory organs may be a means of preventing mosquitoesfrom finding and biting their human victims.

Chemical odorants for use in an olfactometer such as lactic acid orammonia, for example, are available from commercial sources and preparedby standard methods. In some instances, a concentration gradient ofodorant from 0.001 to 100 mg/ml, for example, is used to assess themosquito response. Human sweat for olfaction experiments may becollected from the foreheads or other body parts of human volunteersundergoing physical exercise in a warm, humid environment. The sweat iseither frozen immediately to −20° or allowed to incubate at 37° C. forseveral days. Work from Braks, et al. (referenced above) suggests thatwhile fresh human sweat can be a mild attractant, sweat that has been“aged” is a particularly potent attractant. Other methods for extractingskin odorants include continuous swabbing of human skin with a cottonswab for about 5 minutes or simply inserting a human extremity (e.g., afinger) into the trapping port (see, Dekker, et al., Medical VeterinaryEntomology 16:91-98, 2002, which is incorporated by reference herein).

In addition to blood meal, female as well as male mosquitoes feed onplant nectar as an energy source, which they locate chiefly by visualand chemical cues. Nectar sources do not appear to be as attractive asblood sources, but sugar feeding is usually necessary and more frequentthan blood feeding (see, e.g., Foster & Hancock. J Am Mosquito ControlAssn. 10:288-296, 1994, which is incorporated by reference herein). Assuch, the effects of laser treatment on the ability to locate a nectarsource can also be assessed.

The structural integrity of antennae following laser treatment may beassessed using light microscopy or scanning electron microscopy (see,e.g., Pitts & Zwiebel, Malaria J. 5:26, 2006, which is incorporated byreference herein). For light microscopy, the antennae are hand dissectedfrom cold-anesthetized, laser treated or untreated mosquitoes and placedin 25% sucrose and 0.1% Triton X-100 in water. The antennae are mountedon microscope slides in this solution, covered with a glass coverslip,and sealed with, for example, enamel nail polish. Standard lightmicroscopy at 400× magnification is used to assess the integrity of theantennae.

For scanning electron microscopy, the antennae from either laser treatedor untreated mosquitoes are hand dissected and fixed with 4%paraformaldehyde, 0.1% Triton X-100 in phosphate buffered saline. Theantennae are then dehydrated through a series of alcohol solutions suchas ethanol at 50% to 100% in 10% increments. The heads are furtherextracted through a series of ethanol:hexamethyldisilazane (HMDS)solutions at ratios of 75:25, 50:50, 25:75 and 0:100. The HMDS isremoved and the samples are allowed to dry in a fume hood. Thedesiccated samples are glued onto pin mounts with colloidal silver paintand sputter coated for about 30 seconds with gold-palladium. The samplesare viewed using a standard scanning electron microscope. Alternatively,the antennae are quick frozen in liquid nitrogen and subsequently freezedried to remove any water vapor in preparation for cryo-scanningelectron microscopy at −190° C. In some instances, the head or wholemosquito is used for analysis.

Electroantennography (EAG) is a method for recording electricalpotentials from insect antennae in response to stimuli and can be usedto assess the functional integrity of antennae following treatment withthe laser. EAG records the “slow” changes in potential that are causedby the superposition of simultaneous membrane depolarizations ofnumerous receptor cells in response to stimuli. This approach canprovide information on the olfactory perception of the insect. Anelectroantennogram can be performed by removing the antenna from eitherlaser treated or untreated mosquitoes and inserting wires at either endsof the antenna and amplifying the voltage between. The antenna isexposed to an odorant and any deflections in the electroantennogramwaveform due to sensory response are recorded. Alternatively, a lasertreated or untreated mosquito is left intact and a ground wire or glasselectrode is placed into some part of the body such as the eye, forexample, and a second electrode is attached to the end of the antenna.Alternatively, all or part of a laser treated or untreated mosquito isfixed on the tip of a holder with a conducting electrode gel. The tip ofthe antenna is pushed into a small drop of the same gel associated witha recording electrode (silver wire; see, e.g., Puri, et al., J. Med.Entomol. 43:207-213, 2006, which is incorporated by reference herein).The antenna is exposed to odorant and changes in the electroantennogramwaveform are noted. Using this approach, the normal response to odorantsin untreated mosquitoes can be compared with the response recorded inlaser treated mosquitoes.

To assess whether specific sensilla on the antenna or maxillary palphave been damaged by the laser treatment, odor response at the olfactorysensory level can be done using sensilla recording. The sensilla containolfactory receptor neurons and action potentials of single neurons canbe recorded in situ and the olfactory receptor neurons classifiedaccording to their response to various odorant stimuli. In thistechnique, microelectrodes are inserted into the base of a sensillum andmoved with a micromanipulator to a position at whichelectrophysiological activity can be recorded. The signals are digitizedand observed as spikes of activity. The antenna is exposed to a puff ofodorant and the firing frequency of the neuron is recorded. As above,the normal response to odorants in untreated mosquitoes can be comparedwith the response recorded in laser treated mosquitoes.

The antennae of mosquitoes are also important for sensing the proximityof a potential mate (see Hoy, PNAS 103:16619-16620, 2006; Cator et al.,Science published on line Jan. 8, 2009, both of which are incorporatedby reference herein). More specially, male mosquitoes detect thepresence of nearby female mosquitoes by hearing the female's flighttones using a special organ called the Johnston's organ at the base ofeach antenna. A mosquito detects the particle velocity component of asound field in its immediate vicinity. The antenna, with its fine,flagellar hairs, senses movements of air particles as they are movedabout by incoming acoustic waves. A male mosquito is able to hear anearby female's wing beat frequency (approximately 300-600 Hz, dependingupon the species) and fly off in pursuit. In the case of Aedes aegypti,both male and female mosquitoes are able to adjust the harmonicresonance of their thoracic box to produce a harmonic frequency that isthree times that of the female wing beat (400 Hz) and two times that ofthe male wing beat (600 Hz), converging at a frequency of 1200 Hz at thetime of mating (Cator et al.). In this instance, mate attraction isacoustically driven and involves active modulation by both sexes.

During the mating process, the ability to hear the appropriate flighttones of a nearby female is dependent upon the antennae and associatedJohnston's organ. Likewise, the ability to generate a wing beatfrequency capable of attracting a mate is dependent upon functionalwings. As such, disabling the antennae or wings would potentiallyprevent productive mating.

In general, females emerge from the pupal case ready to mate where astheir male counterpart in many species may require several days to reachsexual maturity. However, in most species, there is a 24-48 hour lagbetween emergence and mating. Mating is not needed for egg developmentand maturation, but in most species eggs can only be deposited wheninsemination has occurred. Female mosquitoes usually mate before takingtheir first blood meal, although in several anophelines, a largepopulation of virgins may blood-feed prior to mating. In Aedes aegypti,mating is accompanied by the transfer of “matrone”, a male hormone whichmakes the female refractory to successive matings and induces bloodhost-seeking behavior. This type of behavioral change is notconsistently noted in An. gambiae. The success of male mating isdetermined by fitness, and may have consequences for the number of timesa male can mate. A number of issues regarding mating behavior have notbeen fully explored or understood including the cues that control maleswarming, male feeding behavior and fitness, female mate-locationbehavior, pre- and post-mating behavior, frequency of multiple-speciesswarming, factors that prevent hybridization of closely related species,and factors that control multiple mating (as outlined by Takken et al.,in “Mosquito mating behaviour”, in Bridging laboratory and fieldresearch for genetic control of disease vectors. pp. 183-188, Ed. G. J.Knols & C. Louis, Springer, Netherlands, 2006, which is incorporated byreference herein).

Male fitness and associated reproductive success may be a function of anindividual's ability to find and exploit a nectar source (see, e.g.,Yuval et al., Ecological Entomology. 19:74-78, 2008, which isincorporated by reference herein). Males tend to swarm at dusk, abehavior that consumes a considerable amount of energy relative toresting behavior. Females enter the swarm of males for mating purposes(see, e.g, Charlwood, et al., J. Vector Ecology 27:178-183, 2003, whichis incorporated by reference herein). Sugar feeding in An. freeborni,for example, takes place during the night at a time after swarming hasconcluded and as such nectar sugars are not immediately available forflight but must be stored in some form. As such, disrupting the abilityto fly or the ability to find or store an energy source will havedeleterious effects on mating success.

Alterations in wing beat frequency in response to laser treatment can beassessed using a particle velocity microphone as described by Cator, etal. (Science Published on line Jan. 8, 2009). Either laser treated oruntreated mosquitoes are tethered to the end of an insect pin. Whensuspended in midair, the mosquitoes initiate bouts of wing-flappingflight. Sound clips from normal and laser treated mosquitoes aredigitized and compared to assess the effects of laser treatment on wingbeat frequency. Alternatively, high speed photography can be used toassess changes in wing function.

Thermal stress may be used to alter the normal embryonic development ofmosquito eggs. Huang, et al. demonstrated that subjecting mosquito eggsto increasing temperatures from 40 to 48° C. reduced the viability ofthe eggs (see, e.g., Huang, et al., Malaria J. 5:87, 2006, which isincorporated by reference herein). Exposure to temperatures of 44-45° C.and higher dramatically decrease the number of eggs that hatched. Assuch, subjecting the female mosquito to laser induced thermal stress mayalso alter the viability of her eggs.

In one set of experiments, female mosquitoes are allowed to blood feedand are subsequently subjected to laser treatment as described herein.Following a recovery period and prior to laying of eggs, the femalemosquitoes are cold-anesthetized and the eggs are dissected out andcounted. The eggs may be further subjected to scanning electronmicroscopy or other forms of microscopy to determine whether treatmentwith the laser has disrupted the structural integrity of the eggs. Forexample, the various stages of ovogenesis in mosquitoes may be assessedusing scanning electron microscopy (Soumare & Ndiaye. Tissue & Cell.37:117-124, 2005, which is incorporated by reference herein).Alternatively, the females are allowed to lay their eggs following lasertreatment. In this instance, the number of eggs laid, the number ofhatched eggs, and the number of viable offspring are compared betweenlaser treated and untreated individuals.

In a second set of experiments, female mosquitoes are subjected to lasertreatment as described herein prior to blood feeding. After bloodfeeding, the females are allowed to lay their eggs and as above, thenumber of eggs laid and the viability of the eggs are determined. Inthese experiments, the number of females that take an offered blood mealmay also be determined in exploring effects on fertility.

Blood feeding is necessary for the process of laying and hatching viableoffspring. Disrupting the ability of the female to access blood meal isanticipated to reduce the number of viable offspring. As noted above,the female uses olfaction to find a blood host. As such, in one set ofexperiments, the blood meal is placed on the other side of a trap portalthrough which the mosquito must pass to access food. The trap portalemits an attracting human odorant such as human sweat or expired carbondioxide. The ability of laser treated females to access the blood mealis recorded as is the number of laid eggs, the number of hatched eggs,and the number of viable offspring.

In general, the effects of laser treatment on male and female fertilitycan be assessed by treating either a population of males or a populationof females with laser energy and allowing the treated individuals of onesex to breed with untreated individuals of the other sex. As above, theoutcome measurement of this assessment is the number of laid eggs, thenumber of hatched eggs, and the number of viable offspring. For thepurposes of this experiment, male and female individuals are treatedwith laser energy prior to mating. Male and female individuals can besexed at the larval stage, allowing for the isolation of single sexpopulations (see, e.g., Emami, et al., J. Vector Borne Dis. 44:245-24,2007, which is incorporated by reference herein). For example, male An.stephensi mosquitoes are identified by a tube-like organ at the 9^(th)abdomen segment as well as two fried egg-shaped structures in theanterior portion of the segment. In female An. stephensi mosquitoes, thetube-like organ is smaller and the fried egg-shaped structures areabsent. Using a light microscope, it is possible to segregate the larvainto separate male and female populations. Alternatively, sexing may bedone following emergence from the pupal stage. Adult male mosquitoes canbe distinguished from adult female mosquitoes in that the males havemore feathery antennae and have mouthparts not suitable for piercingskin. The emerged adults in the single sex populations are subjected tolaser treatment and after recover are allowed to breed with untreatedindividuals of the opposite sex. The number of copulas is observed andrecorded over a specific time frame. In addition, the number of laideggs, hatched eggs, and viable offspring are recorded and may beassessed relative to the number of observed copulations. Similarexperiments can be performed using populations of male and femalemosquitoes that have both been subjected to laser treatment.

Calorespirometry can be used to measure respiration characteristics andenergy metabolism of insects (see, e.g., Acar, et al., Environ. Entomol.30:811-816, 2001; Acar, et al., Environ. Entomol. 33:832-838, 2004, bothof which are incorporated herein by reference). The rates of respiratorymetabolism are commonly reported as the rates of oxygen (O₂) consumptionor carbon dioxide (CO₂) production and may be combined with heatproduction to assess metabolic efficiency. Analysis is done comparingthe response of laser treated and untreated mosquitoes. The analysis canbe done at one specific temperature such as, for example, an ambienttemperature of 27° C. Alternatively, the effects of temperature onmetabolic efficiency of treated and untreated mosquitoes can be assessedby performing the analysis at various temperatures ranging from about 0°C. to about 42° C. In this instance, temperature acts as a stressor.

A differential, scanning, heat conduction calorimeter is used forcalorespirometry (e.g., Hart Scientific model 7707 or calorimetrySciences model 4100, Pleasant Grove, Utah). One or more mosquitoes foranalysis are weighed and placed in a small paper cage within a sampleampoule. The cage is used to limit the mobility of the mosquitoes duringanalysis. The ampoule is supplied with sufficient oxygen to supportaerobic respiration for at least one hour. Heat production is measuredby the calorimeter and is represented as a function of body weight. CO₂production is assessed by measuring extra heat generated over time when0.4 M NaOH is included in the ampoule. The interaction of NaOH with theCO₂ produced by the respiring tissue generates Na₂CO₃ and heat. As such,the difference in heat rate produced by the mosquito sample with andwithout NaOH represents the heat rate caused by CO₂ trapping andconsequently the rate of CO₂ formation. Analysis of heat and CO₂production is performed at various temperatures to assess the effect ofthermal stress on mosquitoes that have been treated with laser energyrelative to untreated controls.

Photonic Dosing Experiments

A series of experiments examining the vulnerability to radiation of An.stephensi has been performed. Dosing experiments began by removing thefood, water, and any other materials from the (floor of the) C2G box.Then the box was moved into the optics room. The mesh holes were looselycovered, and tubing from a CO₂ tank was hooked to the port on the C2Gbox. CO₂ was turned on, with the regulator opened up as wide aspossible, resulting in roughly 50 scfh for a minute or so, until all ofthe mosquitoes were anesthetized. Then the CO₂ flow was turned down to amuch lower level, typically 7-10 scfh.

FIG. 5 is a graph illustrating lethality of various doses of near-IRradiation as a function of energy density. The diode laser, capable ofoutputting up to 30 W of 808 nm light, was manufactured by Coherent,Inc. Optics were used to focus the beam to roughly 5 mm diameter at themosquito. Pulse duration was varied from ˜3 ms up to ˜25 ms, and laseroutput power was varied from ˜15 W up to ˜30 W. Mosquitoes present inthese experiments were predominantly female, although some males mayhave been present in some of the experiments. Subjects were exposed toCO₂ for 8-15 minutes during the experiments. Lethality is measured 24hours after dosing.

FIG. 6 and FIG. 7 are graphs illustrating lethality of various doses ofultraviolet radiation for different power densities and total energiesfor female and male An. stephensi, respectively. The dosing laser forthese experiments was a high power water cooled deep UV laser fromPhotonix, operating at a wavelength of 266 nm. The data underlying thesegraphs are summarized in Table 1.

TABLE 1 Female Male Power Total # Males # Females Survival % Survival %Density Energy @ start @ start 24 hrs 24 hrs (W/cm{circumflex over( )}2) (mJ) 23 11 18% 13% 6.94E+06 16.74 26 27 22% 19% 6.94E+06 8.37 1240 15%  8% 1.78E+07 5.58 34 8  0%  9% 1.78E+07 2.232 35 11 18%  6%6.94E+06 2.232 19 12 25% 11% 2.24E+06 1.488 30 16 81% 43% 3.65E+04 0.28619 16 75% 37% 3.65E+04 3.3 7 24 83% 29% 2.24E+06 1.24 2 22 77% 50%2.24E+06 0.992 8 21 95% 100%  6.38E+05 6.2 8 18 100%  88% 6.38E+05 0.99217 26 88% 65% 6.38E+05 6.2 8 24  0%  0% 6.38E+05 24.8 8 18  0%  0%2.24E+06 24.8 8 17 35%  0% 1.97E+06 6.696 13 21 90% 77% 1.29E+05 7.75 1320 95% 92% 1.29E+05 1.395 8 12 100%  100%  6.38E+04 6.2248 7 20 100% 100%  6.94E+05 4.464 4 21 100%  100%  2.24E+05 8.06 4 17 100%  100% 2.24E+05 1.488 4 29 97% 75% 6.94E+05 1.2834 12 17 100%  92% 6.94E+050.4464 30 8 25% 10% 6.94E+06 19.53 23 17  0% 13% 6.94E+06 9.486 22 2241%  5% 6.94E+06 4.464 22 19 89% 91% 0.00E+00 0 22 19 68% 45% 6.94E+061.674 8 29 10% 25% 6.94E+06 13.95 16 29 10%  0% 6.94E+06 5.022 10 29 90%30% 2.24E+06 2.232 22 24 38%  9% 2.24E+06 6.944 14 23 22% 21% 2.24E+0614.88

It will be seen that each graph includes two regimes: at lower powerdensities, survival fraction is primarily a function of total energydeposited in the insect's body. At higher power densities, the energyrequired to kill an insect decreases, and survival fraction is primarilya function of power density. It is believed that this is due to theoptical saturation of absorbing molecules (sometimes described asphotobleaching) in the insect's exoskeleton and other surface layers,and the consequent penetration of light into interior tissues which aresubject to photochemical damage, particularly of active DNA.

The experiments reported in FIG. 5, FIG. 6, and FIG. 7 use 24-hoursurvival fraction of mosquito population as a figure-of-merit. In someembodiments, it may be sufficient to disable, rather than kill,mosquitoes or other targets, as discussed elsewhere herein. Further, thelife cycle of malaria requires a period of approximately 11-14 daysbetween infection of a mosquito and transmission to a human host. Thus,it may be possible to substantially impact malaria rates by achieving asuitably low 10-day survival fraction, which may require differentenergies or power densities than those shown in the reported data.Finally, it is unknown to what extent anesthetization and handling mayaffect energies or power densities required to affect mosquitoes.Experiments similar to those reported in FIG. 5, FIG. 6, and FIG. 7 butusing the tracking and targeting systems described herein may providefurther information about suitable systems for disabling mosquitoes orother pests.

Trap Validation

Systems such as those shown and described herein may be used to measurethe efficacy of traps and to identify the most reliable methods ofmonitoring insect populations. The World Health Organization haspublished “Dengue: Guidelines For Diagnosis, Treatment, Prevention AndControl,” a copy of which is included herewith and which is incorporatedby reference herein, describing in Section 5.2.2 methods ofentomological surveillance of dengue vectors (in particular, Aedesaegypti). Current methods include sampling larvae and pupae,pupal/demographic surveys, sampling the adult mosquito population,landing collections, resting collections, sticky trap collections,oviposition traps, and larvitraps. Some of these methods are expensiveand involve potential ethical concerns (e.g., landing collections, whichmay involve human contact with possibly-infected mosquitoes), and it isnot well understood how well any of these methods correlate with adultmosquito populations. The present invention will permit inexpensivemethods such as sticky traps and larvitraps to be compared with adultpopulations to determine whether these methods provide adequate measuresof mosquito populations.

Prophetic Example 1: Surveillance of Mosquitoes with a Photonic SystemVersus an Ovitrap

A photonic system is used to identify and enumerate the number ofmosquitoes flying over and around an ovitrap device. The efficacy andaccuracy of the photonic system versus the ovitrap device in monitoringthe number of mosquitoes infesting a site are compared. The photonicsystem includes an imager, an illumination source, a retroreflector anda processor to locate and identify mosquitoes. The ovitrap comprises ajar containing water, a mosquito attractant, and wooden paddles tocollect and count eggs deposited by females traversing the site. Theprevalence of a mosquito vector, Aedes aegypti, is measured using aphotonic system and ovitraps.

An oviposition trap (aka ovitrap) is used to detect the presence ofmosquitoes and to monitor the density of mosquitoes in a village. Eachovitrap includes a 350 mL cup painted black with seed germination papercovering the inside of the cup, and with approximately 175 mL of hayinfusion to attract mosquitoes. Methods and materials to make enhancedovitraps are described (see e.g., Polson et al., Dengue Bulletin 26:178-184, 2002 which is incorporated herein by reference). The ovitrapsare placed approximately one meter off the ground in a shelteredlocation to avoid rainfall and sun and left for 48 hours, and then theseed germination paper is removed and sent to a lab for mosquito eggcounting which is done manually with the aid of magnification. Speciesidentification requires rearing larvae from the eggs. The ovitrap isreset with fresh germination paper and hay infusion fluid for another 48hours, and the process is repeated for approximately 4 weeks. To samplea rural village or a city, 50-262 ovitraps may be required (see e.g.,Polson et al., Ibid., and Regis et al., PLoS ONE 8: e67682doi:10.1371/journal.pone.0067682 which are incorporated herein byreference). The surveillance data obtained from the ovitraps mayinclude: The percentage of traps with mosquito eggs present; the numberof mosquito eggs per ovitrap and the corresponding locations of thepositive traps. For example, ovitraps with hay infusion placed in avillage outside of Phnom Penh, Cambodia detected mosquito eggs in 9%-67%of outdoor traps over thirteen trap collections, and a mean number of4-23 eggs per trap over thirteen collections (see e.g., Polson et al.,Ibid.).

The photonic system employs a high speed CMOS camera, a retroreflectorscreen, an illumination source and a processor to acquire and analyzethe images obtained by the system and to determine biological parametersfrom the mosquito images. For example, the camera may be a Phantom Flexavailable from Vision Research, Wayne, N.J. which has a variable shutterspeed and frame rates exceeding 10,000 frames/second (see e.g.,Datasheet for Phantom Flex camera, which is incorporated herein byreference). Image acquisition and image processing software may beprovided with the camera or separately. Alternative computer programs totrack and record the flight path of flying insects are described (seee.g., Spitzen et al., in Proceedings of Measuring Behavior 2008,Maastricht, The Netherlands, Aug. 26-29, 2008 eds. Spink et al., whichis incorporated herein by reference). The photonic system also includesan illumination source and a retroreflector to efficiently reflect lightfrom the light source back to the camera (see FIG. 1). For example alight emitting diode and a reflector surface including retroreflectorfabric such as SCOTCHLIGHT™ Silver Industrial Wash Fabric 9910,available from 3M Corp., may be used to backlight insects as they flyacross the camera's field of view. Microcircuitry on the device analyzesthe image data to identify, locate, and enumerate mosquitoes enteringthe field of view over a defined period (e.g., 48 hours). For examplethe identity of a flying insect may be determined by the varyingamplitude of a specific wavelength of light reflected from the insect'sbeating wings as described above. Methods to locate and track mosquitoesin flight based upon computerized analysis of video camera images aredescribed (see e.g., Spitzen et al., Ibid.) Moreover, processing of thevideo data for mosquitoes allow determination of multiple parametersincluding species and sex of flying mosquitoes. For example male andfemale Aedes aegypti and Aedes triseriatus mosquitoes may be identifiedand differentiated based on digital recordings of light reflecting offthe mosquitoes in flight. Spectral patterns corresponding to wingbeatfrequencies may be analyzed to obtain a plot of frequency versusamplitude, and computer methods are used to identify species and sex ofclosely related mosquito species (see e.g., Moore, J. Insect Behavior,4(3):391-396 (2005). Robertson, et al., J. Amer. Mosquito ControlAssoc., 18(4):316-320 (2002); and “An Automated Flying-Insect DetectionSystem,” NASA Technical Briefs, SSC-00192 (2007), available at<www.techbriefs.com/content/view/2187/34/> all of which are incorporatedherein by reference). The photonic system is constructed as arectangular enclosure which is placed directly above an enhanced CDCovitrap to allow comparison of the two systems for monitoringmosquitoes.

In comparison, the photonic systems placed over each ovitrap monitor theairspace over the trap and detect and record image data for eachmosquito, male or female regardless of species and egg-laying status.The imaging data is automatically processed to determine the sex andspecies, as well as other biological parameters of any and allmosquitoes which fly through the airspace over the ovitrap. Data on thesex, species and numbers of mosquitoes detected at a specific site overa selected period of time (e.g., 48 hours) is transmitted to acentralized computer or database immediately. In contrast to ovitrapsystems, the counting and reporting of mosquitoes is automated anddependent on algorithms for image analysis. Moreover, the detection ofadult mosquitoes eliminates indirect estimation of gravid females basedon egg counting. By comparing the data generated by the ovitrap systemwith that measured by the photonic system, researchers may gain insightinto the accuracy and efficacy of the ovitrap system.

Prophetic Example 2: Comparison of a Photonic System to Funnel Traps forMeasuring Mosquito Infestation in Wells or Water Containers

A photonic system is used to identify and enumerate the number of adultmosquitoes flying over and around subterranean wells and watercontainers. The photonic system is compared to funnel traps for efficacyand accuracy in monitoring the number of mosquitoes infesting a site.The photonic system includes an imager, an illumination source, aretroreflector and a processor to locate, identify and characterizemosquitoes in flight. The funnel trap is a floating trap which catchesmosquito larvae as they swim to the surface of the well or watercontainer. Larvae counts are done manually to discriminate mosquitolarvae from other insects. The density of Anopheles mosquitoes ismeasured in field tests at subterranean wells with known mosquitoinfestations using a photonic system versus funnel traps.

Funnel traps are tested in water wells to compare their efficacy andaccuracy in monitoring mosquito infestations. Methods and materials toconstruct and test funnel traps are described (see e.g., Russell et al.,J. Med. Entomol. 36: 851-855, 1999 which is incorporated herein byreference). For example, a funnel trap is constructed from a plasticcontainer with a plastic funnel inserted in the lid of the container.The container serves as a reservoir to collect mosquito larvae whichswim upward through the funnel into the reservoir and are trapped. Thefunnel trap is approximately 180 mm long and floats with the funnelmouth (185 mm diameter) facing the bottom of the well. Field tests aredone on 100 cm diameter wells. A funnel trap is set on each wellovernight and mosquito larvae are counted manually after approximately12 hours. Funnel traps sample approximately 20% of the larvae introducedin a well in a single 12 hour sampling period. In field tests theabsolute number of larvae introduced in the traps is predicted with84-97% accuracy with coefficients of variation between 14-39% whenreplicate samplings are done. However, single samplings only allowqualitative prediction of low, medium and high densities of larvae.Funnel traps are less efficient at sampling different mosquitoes. Forexample, Aedes larvae are sampled more efficiently than Culis larvae(e.g., 1.7-2.3 times more efficient) likely due to differing swimmingbehavior of the larvae. Also some stages of mosquito development aresampled less efficiently by funnel traps. For example, 1^(st) and 2^(nd)instar and pupae are trapped at lower efficiency. Funnel trap samplingefficacy also varies with well diameter and thus complicates predictionof larval population size. See e.g., Russel et al., Ibid.

The photonic system includes imagers, illumination sources,retroreflectors and processors to analyze spectral and image data tolocate, track, identify and characterize mosquitoes flying into thefield of view. A high speed camera, capable of 1,000 frames per secondwith high resolution, and with variable shutter speeds (see e.g.,Datasheet for Phantom Flex camera which is incorporated herein byreference) is used to detect and characterize mosquitoes at differentshutter speeds. For example, initial detection and tracking ofmosquitoes entering the field of view may be done at approximately 500frames per second and then imaging of wingbeat frequencies on thetargeted mosquito may be done at 5,000 frames/second. Mosquito wingbeatfrequencies and associated harmonics may range between 500 and 2000cycles per second (see e.g., Moore, J. Insect Behavior, 4(3):391-396(1991) which is incorporated herein by reference). The photonic systemmay include illumination source(s) (e.g., light emitting diodes) andretroreflectors to backlight mosquitoes entering the field of view. Thephotonic system may be bounded by rectangular or cylindrical supportswith imagers, illumination sources, lasers, photodiodes andretroreflectors placed as indicated in FIG. 1. Processors analyzeimaging data and spectral data to locate, identify and track mosquitoesentering the field of view (see e.g., Spitzen et al., Ibid.), moreover,processors may initiate programmed changes in the photonic system. Forexample, identification of a mosquito based on imaging with the highspeed camera at 500 fps may trigger tracking and targeting with a pulsedlaser at 1180 nm to detect hemozoin indicative of malarial infection.The system may also estimate malarial status on the basis of mosquitobehavior, such as changes in flight paths, speed, host-seeking behavior,altitude, or time of day of mosquito activity. See, e.g., Cator et al.,Trends in Parasitol. 28(11):466-470 (2012), Lacroix et al., PLOS Biol.3(9):1590-1593 (2005), Smallegange et al., PLoS ONE 8(5):1-3 (May 2013),all of which are incorporated herein by reference. The photonic systemmay be implemented with a rectangular boundary and installed immediatelyabove water wells containing funnel traps.

Photonic systems are installed over approximately 12 wells containing 1funnel trap each. The photonic systems monitor the airspace over thewells and automatically report the number, species, sex, and probableparasite status (e.g., Plasmodium positive or negative) of mosquitoesthat enter the field of view. For example, over a period of 48 hoursemergent mosquitoes from the well and all other mosquitoes flying intothe field of view are counted and characterized. The data areautomatically transmitted to a central computer for analysis, e.g.,comparison to funnel trap data. After 48 hours the funnel traps areretrieved from the wells and mosquito larvae are visually identified andcounted. The data are manually entered into a computer and compared tothe number of mosquitoes flying over the corresponding wells. Thecorrelation coefficient for the number of mosquito larvae and the numberof flying mosquitoes detected in the wells is calculated. A photonicsystem may provide increased accuracy relative to a funnel trap sincethe determination of mosquito species, sex and other characteristicsconfirms the identification; also the continuous surveillance of theairspace over the well is preferable to the coverage of funnel filters(e.g., 2.4% of a 1.2 m diameter well). Moreover the photonic system isnot subject to the variation in behavior of different mosquito species(e.g., Aedes, Culis, Anopheles) and different larval stages (e.g., seeabove and Russel et al., Ibid.) which complicate the funnel trap system.Finally, the identification of flying mosquitoes infected with a malariaagent, Plasmodium, is important information obtained with a photonicsystem that is not available from analysis of mosquito larvae.

Prophetic Example 3: Comparison of a Photonic Detection System with aHuman Landing Collection Method to Monitor Mosquitoes

A photonic system is compared to a human landing catch (HLC) method tomonitor mosquito density in an African village. The photonic system isconstructed to detect, count and characterize any mosquitoes crossing aperimeter established around selected houses in the village. Individualsin each house are trained to sample host seeking mosquitoes using a HLCmethod. The sensitivity and efficacy of each method for monitoringmultiple species of mosquito is compared.

The photonic system is constructed to image mosquitoes in flight andprocess the imaging data to identify, enumerate, and characterize themosquitoes and to report information on the mosquitoes to a systemcomputer. The photonic system is set up to monitor a perimetersurrounding each of three houses selected for the study. Support postsapproximately 20 cm×20 cm×500 cm high are set approximately 100 m apartto define a perimeter around each house (see FIG. 2). Two high speedcameras (see e.g., Datasheet for Phantom Flex camera, which isincorporated herein by reference) are placed facing each other on eachside of the perimeter to create a photonic fence. The fields of view oneach side of the perimeter are approximately 500 cm high, 100 m long and20 cm thick. Each support post is covered with retroreflective fabric(such as SCOTCHLITE™ 9100 from 3M Corp. in St. Paul, Minn.) to providebacklighting to any mosquitoes crossing the field of view, i.e., thephotonic fence. The photonic system may also have a laser light sourceand a photon detector incorporated on each side of the perimeter. Forexample, a Ti:Sapphire laser producing laser pulses at 1180 nm and aphoton detection system may be used to detect hemozoin, a pigmentassociated with malarial parasites, e.g., Plasmodium (see e.g., Belisleet al., Biophys J. 94(4): L26-L28, Feb. 15, 2008; doi:10.1529/biophysj.107.125443 which is incorporated herein by reference).The photonic system established on the perimeter also includesprocessors, circuitry and programming to identify, locate, count anddetermine biological properties of mosquitoes which cross the perimeter.For example spectral patterns corresponding to wingbeat frequencies maybe analyzed to obtain a plot of frequency versus amplitude, and computermethods are used to identify species and sex of closely related mosquitospecies (see e.g., Moore, J. Insect Behavior, 4(3):391-396 (2005).Robertson, et al., J Amer. Mosquito Control Assoc., 18(4):316-320(2002); and “An Automated Flying-Insect Detection System,” NASATechnical Briefs, SSC-00192 (2007), available at<www.techbriefs.com/content/view/2187/34/> all of which are incorporatedherein by reference). Detailed information on all mosquitoes crossingthe photonic fence is automatically transmitted to a central computer inreal time to create a record of mosquitoes observed every 12 hours(between 7 pm and 7 am) for 7 days or longer. Importantly, informationon the number, species, sex, feeding status, malarial infection andmating status of the mosquitoes is reported.

A human landing catch (HLC) method is established at each of the houseswith a photonic fence system and the data on mosquitoes detected by bothsystems is compared. To collect HLC data an adult male collector exposeshis lower limbs and collects mosquitoes when they land on his legs withan aspirator. The catcher collects mosquitoes for 45 minutes/hour andrests for 15 minutes. Mosquito collections are done nightly between 7 pmand 7 am for 7 days or longer. The HLC catcher collects at an indoorsite and an outdoor site within the perimeter of the photonic fence.Aspirated mosquitoes are processed to identify sex and species bymorphology with a dissecting microscope. Abdominal status is classifiedas fed, unfed, gravid or partly gravid. For example, male and femaleAnopheles are sorted, and females are analyzed for malarial(circumsporozite) proteins using an ELISA assay. Also polymerase chainreaction (PCR) is used to identify mosquito subspecies. Methods andmaterials to conduct HLC, aspirate and process mosquitoes are described(see e.g., Sikaala et al., Parasites and Vectors 6:91, 2013 online at:<www.parasitesandvectors.com/content/6/1/91> which is incorporatedherein by reference). Data on mosquitoes collected with HLC for 12 hourseach night over 7 days is entered into a centralized computer andcompared to photonic fence data collected over the same time frame.

The HLC method and the photonic fence are compared with respect to: theabsolute number of mosquitoes detected for each species, the number offemale mosquitoes, the number of infected mosquitoes (Plasmodium), thenumber of fed vs. unfed mosquitoes, and the mating status of the femalemosquitoes. The efficacy and accuracy of the photonic system versus theHLC may depend on the diligence and stamina of the HLC catchers whocollect 12 hours per night for 7 days or more. Also the risk ofinfection by Plasmodium and other vector-borne diseases is a majordrawback of HLC.

In a general sense, those skilled in the art will recognize that thevarious aspects described herein which can be implemented, individuallyor collectively, by a wide range of hardware, software, firmware, or anycombination thereof can be viewed as being composed of various types of“electrical circuitry.” Consequently, as used herein, “electricalcircuitry” includes, but is not limited to, electrical circuitry havingat least one discrete electrical circuit, electrical circuitry having atleast one integrated circuit, electrical circuitry having at least oneapplication specific integrated circuit, electrical circuitry forming ageneral purpose computing device configured by a computer program (e.g.,a general purpose computer configured by a computer program which atleast partially carries out processes or devices described herein, or amicroprocessor configured by a computer program which at least partiallycarries out processes or devices described herein), electrical circuitryforming a memory device (e.g., forms of memory (e.g., random access,flash, read only, etc.)), or electrical circuitry forming acommunications device (e.g., a modem, communications switch,optical-electrical equipment, etc.). Those having skill in the art willrecognize that the subject matter described herein may be implemented inan analog or digital fashion or some combination thereof.

Those skilled in the art will recognize that at least a portion of thedevices or processes described herein can be integrated into an imageprocessing system. Those having skill in the art will recognize that atypical image processing system generally includes one or more of asystem unit housing, a video display device, memory such as volatile ornon-volatile memory, processors such as microprocessors or digitalsignal processors, computational entities such as operating systems,drivers, applications programs, one or more interaction devices (e.g., atouch pad, a touch screen, an antenna, etc.), control systems includingfeedback loops and control motors (e.g., feedback for sensing lensposition or velocity; control motors for moving/distorting lenses togive desired focuses). An image processing system may be implementedutilizing suitable commercially available components, such as thosetypically found in digital still systems or digital motion systems.

Those skilled in the art will recognize that at least a portion of thedevices or processes described herein can be integrated into a dataprocessing system. Those having skill in the art will recognize that adata processing system generally includes one or more of a system unithousing, a video display device, memory such as volatile or non-volatilememory, processors such as microprocessors or digital signal processors,computational entities such as operating systems, drivers, graphicaluser interfaces, and applications programs, one or more interactiondevices (e.g., a touch pad, a touch screen, an antenna, etc.), orcontrol systems including feedback loops and control motors (e.g.,feedback for sensing position or velocity; control motors for moving oradjusting components or quantities). A data processing system may beimplemented utilizing suitable commercially available components, suchas those typically found in data computing/communication or networkcomputing/communication systems.

In some implementations described herein, logic and similarimplementations may include software or other control structures.Electronic circuitry, for example, may have one or more paths ofelectrical current constructed and arranged to implement variousfunctions as described herein. In some implementations, one or moremedia may be configured to bear a device-detectable implementation whensuch media hold or transmit device-detectable instructions operable toperform as described herein. In some variants, for example,implementations may include an update or modification of existingsoftware or firmware, or of gate arrays or programmable hardware, suchas by performing a reception of or a transmission of one or moreinstructions in relation to one or more operations described herein.Alternatively or additionally, in some variants, an implementation mayinclude special-purpose hardware, software, firmware components, orgeneral-purpose components executing or otherwise invokingspecial-purpose components. Specifications or other implementations maybe transmitted by one or more instances of tangible transmission mediaas described herein, optionally by packet transmission or otherwise bypassing through distributed media at various times.

Alternatively or additionally, implementations may include executing aspecial-purpose instruction sequence or invoking circuitry for enabling,triggering, coordinating, requesting, or otherwise causing one or moreoccurrences of virtually any functional operations described herein. Insome variants, operational or other logical descriptions herein may beexpressed as source code and compiled or otherwise invoked as anexecutable instruction sequence. In some contexts, for example,implementations may be provided, in whole or in part, by source code,such as C++, or other code sequences. In other implementations, sourceor other code implementation, using commercially available or techniquesin the art, may be compiled/implemented/translated/converted into ahigh-level descriptor language (e.g., initially implementing describedtechnologies in C or C++ programming language and thereafter convertingthe programming language implementation into a logic-synthesizablelanguage implementation, a hardware description language implementation,a hardware design simulation implementation, or other such similarmode(s) of expression). For example, some or all of a logical expression(e.g., computer programming language implementation) may be manifestedas a Verilog-type hardware description (e.g., via Hardware DescriptionLanguage (HDL) or Very High Speed Integrated Circuit Hardware DescriptorLanguage (VHDL)) or other circuitry model which may then be used tocreate a physical implementation having hardware (e.g., an ApplicationSpecific Integrated Circuit). Those skilled in the art will recognizehow to obtain, configure, and optimize suitable transmission orcomputational elements, material supplies, actuators, or otherstructures in light of these teachings.

In one embodiment, several portions of the subject matter describedherein may be implemented via Application Specific Integrated Circuits(ASICs), Field Programmable Gate Arrays (FPGAs), digital signalprocessors (DSPs), or other integrated formats. However, those skilledin the art will recognize that some aspects of the embodiments disclosedherein, in whole or in part, can be equivalently implemented inintegrated circuits, as one or more computer programs running on one ormore computers (e.g., as one or more programs running on one or morecomputer systems), as one or more programs running on one or moreprocessors (e.g., as one or more programs running on one or moremicroprocessors), as firmware, or as virtually any combination thereof,and that designing the circuitry or writing the code for the softwareand or firmware would be well within the skill of one of skill in theart in light of this disclosure. In addition, those skilled in the artwill appreciate that the mechanisms of the subject matter describedherein are capable of being distributed as a program product in avariety of forms, and that an illustrative embodiment of the subjectmatter described herein applies regardless of the particular type ofsignal bearing medium used to actually carry out the distribution.Examples of a signal bearing medium include, but are not limited to, thefollowing: a recordable type medium such as a floppy disk, a hard diskdrive, a Compact Disc (CD), a Digital Video Disk (DVD), a digital tape,a computer memory, etc.; and a transmission type medium such as adigital or an analog communication medium (e.g., a fiber optic cable, awaveguide, a wired communications link, a wireless communication link(e.g., transmitter, receiver, transmission logic, reception logic,etc.), etc.).

It will be understood that, in general, terms used herein, andespecially in the appended claims, are generally intended as “open”terms (e.g., the term “including” should be interpreted as “includingbut not limited to,” the term “having” should be interpreted as “havingat least,” the term “includes” should be interpreted as “includes but isnot limited to,” etc.). It will be further understood that if a specificnumber of an introduced claim recitation is intended, such an intentwill be explicitly recited in the claim, and in the absence of suchrecitation no such intent is present. For example, as an aid tounderstanding, the following appended claims may contain usage ofintroductory phrases such as “at least one” or “one or more” tointroduce claim recitations. However, the use of such phrases should notbe construed to imply that the introduction of a claim recitation by theindefinite articles “a” or “an” limits any particular claim containingsuch introduced claim recitation to inventions containing only one suchrecitation, even when the same claim includes the introductory phrases“one or more” or “at least one” and indefinite articles such as “a” or“an” (e.g., “an imager” should typically be interpreted to mean “atleast one imager”); the same holds true for the use of definite articlesused to introduce claim recitations. In addition, even if a specificnumber of an introduced claim recitation is explicitly recited, it willbe recognized that such recitation should typically be interpreted tomean at least the recited number (e.g., the bare recitation of “twoimages,” or “a plurality of images,” without other modifiers, typicallymeans at least two images). Furthermore, in those instances where aphrase such as “at least one of A, B, and C,” “at least one of A, B, orC,” or “an [item] selected from the group consisting of A, B, and C,” isused, in general such a construction is intended to be disjunctive(e.g., any of these phrases would include but not be limited to systemsthat have A alone, B alone, C alone, A and B together, A and C together,B and C together, or A, B, and C together, and may further include morethan one of A, B, or C, such as A₁, A₂, and C together, A, B₁, B₂, C₁,and C₂ together, or B₁ and B₂ together). It will be further understoodthat virtually any disjunctive word or phrase presenting two or morealternative terms, whether in the description, claims, or drawings,should be understood to contemplate the possibilities of including oneof the terms, either of the terms, or both terms. For example, thephrase “A or B” will be understood to include the possibilities of “A”or “B” or “A and B.”

While various aspects and embodiments have been disclosed herein, otheraspects and embodiments will be apparent to those skilled in the art.The various aspects and embodiments disclosed herein are for purposes ofillustration and are not intended to be limiting, with the true scopeand spirit being indicated by the following claims.

What is claimed is:
 1. A system for tracking organisms, comprising: animager having an image resolution and a field of view; a backlightsource configured to be placed in the field of view of the imager; and aprocessor configured to analyze one or more images captured by theimager including at least a portion of the backlight source, theprocessor being configured to identify a biological property of anairborne organism in the field of view of the imager; and a physicaltrap configured to physically capture at least one organism, wherein thesystem is configured to use the identified biological property tomeasure an efficacy of the physical trap.
 2. The system of claim 1,wherein the identified biological property is selected from the groupconsisting of genus, species, sex, mating status, gravidity, feedingstatus, age, and health status.
 3. The system of claim 1, whereinmeasuring the efficacy of the physical trap includes comparing a numberof organisms in the trap with a number of airborne organisms identifiedby the processor.
 4. The system of claim 3, wherein comparing the numberof organisms in the trap with the number of airborne organismsidentified by the processor includes comparing numbers collected duringthe same time interval.
 5. The system of claim 3, wherein comparing thenumber of organisms in the trap with the number of airborne organismsidentified by the processor includes comparing numbers collected duringdifferent time intervals.
 6. The system of claim 1, wherein the field ofview of the imager includes the trap interior.
 7. The system of claim 1,wherein the field of view of the imager includes a volume exterior tothe trap.
 8. The system of claim 1, wherein the airborne organism is aninsect.
 9. The system of claim 1, wherein the airborne organism is amosquito.
 10. The system of claim 1, wherein the airborne organism is apsyllid.
 11. The system of claim 1, wherein the backlight sourceincludes a retroreflector.
 12. The system of claim 1, wherein thebacklight source is a retroreflector.
 13. The system of claim 1, whereinthe trap is configured to capture flying organisms.
 14. The system ofclaim 1, wherein the trap is configured to capture immature individualsof a species that is capable of flight at maturity.
 15. A method ofdetermining efficacy of a trap for airborne organisms, comprising:monitoring a population of airborne organisms to determine a populationof airborne organisms in a monitored space by: acquiring an image havingan image resolution from an imager, the imager having a backlight sourcein its field of view, the field of view including the monitored space;determining that the image includes an organism; and determining abiological property of the organism; determining a number of organismsof a species capable of flight captured by a trap; and comparing thenumber of captured organisms with the determined population of airborneorganisms.
 16. The method of claim 15, wherein the biological propertyof the organisms is selected from the group consisting of genus,species, sex, mating status, gravidity, feeding status, age, and healthstatus.
 17. The method of claim 15, wherein comparing the number ofcaptured organisms with the determined population of airborne organismsincludes comparing only those organisms having a selected biologicalproperty.
 18. The method of claim 15, wherein comparing the number ofcaptured organisms with the determined population of airborne organismsincludes determining a first fraction of the population of airborneorganisms having a selected biological property and determining a secondfraction of the captured organisms having the selected biologicalproperty, and further includes comparing the first fraction and thesecond fraction.
 19. The method of claim 15, wherein the airborneorganism is an insect.
 20. The method of claim 15, wherein the airborneorganism is a mosquito.
 21. The method of claim 15, wherein the airborneorganism is a psyllid.
 22. The method of claim 15, wherein the backlightsource includes a retroreflector.
 23. The method of claim 15, whereinthe backlight source is a retroreflector.
 24. The method of claim 15,wherein the trap is configured to capture flying organisms.
 25. Themethod of claim 15, wherein the trap is configured to capture immatureindividuals of a species that is capable of flight at maturity.